Fluorescent molecularly imprinted polymer thin films for specific protein detection prepared with dansyl ethylenediamine-conjugated O-acryloyl l-hydroxyproline

• Published in: Biosensors & bioelectronics 2013-10, Vol.48, p.113-119
• Main Authors: Inoue, Yuki, Kuwahara, Atsushi, Ohmori, Kohei, Sunayama, Hirobumi, Ooya, Tooru, Takeuchi, Toshifumi
• Format: Article
• Language: English
• Subjects: Animals; Biological and medical sciences; Biosensing Techniques - methods; Biotechnology; Cattle; Dansyl Compounds - chemistry; Ethylenediamines - chemistry; Fluorescent detection; Fluorescent Dyes - chemistry; Fundamental and applied biological sciences. Psychology; Humans; Hydroxyproline - chemistry; Molecular Imprinting; Molecular recognition; Molecularly imprinted polymers; Polymers - chemistry; Protein sensing; Sensitivity and Specificity; Serum Albumin, Bovine - analysis; Serum Albumin, Bovine - isolation & purification; Spectrometry, Fluorescence - methods; Synthetic receptor
• ISSN: 0956-5663; 1873-4235
• DOI: 10.1016/j.bios.2013.03.005

Protein-imprinted polymers, capable of specific transduction of protein binding events into fluorescent signal change, were designed and synthesized by using dansyl ethylenediamine-conjugated O-acryloyl l-hydroxyproline (Hyp-En-Dans). Human serum albumin (HSA) was used as a model target protein and HSA-imprinted polymers (HSA-IP) were prepared on glass substrates. Specific fluorescence change was observed for HSA binding on the imprinted polymer thin film, whereas a weaker response was observed for other proteins, including bovine serum albumin, chymotrypsin, lysozyme, and avidin. The binding specificity was found to derive from the rigid structure of the hydrogen-bondable pyrrolidine moiety. Compared with SPR measurements, the non-specific binding caused by the polymer matrix and/or randomly located fluorescent monomer residues that did not compose specific binding sites did not contribute to the observed fluorescence change. These results revealed that the proposed protein-imprinting technique using Hyp-En-Dans could provide a highly selective protein-sensing platform, in which only specific binding events would be detected by fluorescent measurements. •Protein-imprinted polymers prepared can transduce binding events into optical change.•A new functional monomer Hyp-En-Dans was designed and synthesized for this purpose.•The binding specificity was derived from the rigid structure of the pyrrolidine moiety.•The non-specific binding did not contribute to the observed fluorescence change.•The proposed imprinting technique could provide a highly selective sensing platform.